Methylphenidate-triggered ROS generation promotes caveolae-mediated transcytosis via Rac1 signaling and c-Src-dependent caveolin-1 phosphorylation in human brain endothelial cells

• Tipo: Artigo em Revista Científica Internacional
• Data: 2016

Autores

Coelho-Santos, V (Autor) Outra; Socodato, R (Autor) Outra; Portugal, C (Autor) Outra; Leitão, RA (Autor) Outra; Rito, M (Autor) Outra; Barbosa, M (Autor) Outra; Couraud, P (Autor) Outra; Romero, IA (Autor) Outra; Weksler, B (Autor) Outra; Minshall, RD (Autor) Outra; Fontes-Ribeiro, C (Autor) Outra; Summavielle, T (Autor) Outra; Relvas, JB (Autor) ICBAS; Silva, AP (Autor) Outra

Revista

Título: Cellular and Molecular Life Sciences

Páginas: 1-16

ISSN: 1420-682X

Editora: Springer Nature

Indexação

Scopus - 0 Citações

Outras Informações

ID Authenticus: P-00K-YBS

DOI: 10.1007/s00018-016-2301-3

Abstract (EN): Methylphenidate (MPH) is an amphetamine-like stimulant commonly prescribed for attention deficit hyperactivity disorder. Despite its widespread use, the cellular/molecular effects of MPH remain elusive. Here, we report a novel direct role of MPH on the regulation of macromolecular flux through human brain endothelial cells (ECs). MPH significantly increased caveolae-mediated transcytosis of horseradish peroxidase through ECs without affecting paracellular permeability. Using FRET-based live cell imaging, together with pharmacological inhibitors and lentiviral-mediated shRNA knockdown, we demonstrate that MPH promoted ROS generation via activation of Rac1-dependent NADPH oxidase (NOX) and c-Src activation at the plasma membrane. c-Src in turn was shown to mediate the phosphorylation of caveolin-1 (Cav1) on Tyr14 leading to enhanced caveolae formation and transendothelial transport. Accordingly, the inhibition of Cav1 phosphorylation by overexpression of a phosphodefective Cav1Y14F mutant or knocking down Cav1 expression abrogated MPH-induced transcytosis. In addition, both vitamin C and inhibition of NOX blocked MPH-triggered vesicular transport. This study, therefore, identifies Rac1/NOX/c-Src-dependent signaling in MPH-induced increase in transendothelial permeability of brain endothelial cell monolayers via caveolae-mediated transcytosis. © 2016 Springer International Publishing

Idioma: Inglês

Tipo (Avaliação Docente): Científica